Transcript
Page 1: Next Generation Recombinant Protein Manufacturing

Next Generation Processes: What Model Works Best to Manufacture Recombinant

Proteins in Asia?

Thomas Jung, M.S.Vice President, Business Development

KBI Biopharma Inc.

BioPharma Asia 2017Suntec Convention

CenterSingapore

March 22, 2017

Page 2: Next Generation Recombinant Protein Manufacturing

Internal Manufacturing vs CMO

Page 3: Next Generation Recombinant Protein Manufacturing

Internal vs CMO Considerations

MoleculeIndication

Target Market(s)Manufacturing Yield

Facilities CostAccess to Expertise

Location of CMO

Page 4: Next Generation Recombinant Protein Manufacturing

Type of Molecule• Innovator vs biosimilar• Therapeutic antibody, recombinant protein,

antigen, peptide• Stage of development: preclinical, early

clinical, late stage, commercial• Indication• Route of administration• Dose: known or estimated• Dosage will influence manufacturing

demand.

Page 5: Next Generation Recombinant Protein Manufacturing

Target Market(s)• Selected country or countries, region, or

global?• Regulatory requirements may influence

manufacturing strategy.• Clinical / commercial drug substance / drug

product requirements.• Market scope will influence DS / DP

requirements and manufacturing demand.

Page 6: Next Generation Recombinant Protein Manufacturing

Manufacturing Yield• Production clone developed? • RCB Available?• Yield of current cell line adequate?• Consider development of higher yielding cell

line.• Higher yielding cell line reduces

manufacturing scale.• Higher productivity cell lines / more costly

licensing terms.

Page 7: Next Generation Recombinant Protein Manufacturing

Internal Manufacturing Considerations

• Costs• Facility cost• Equipment cost• Development and manufacturing personnel cost• Quality systems cost

• Development Strategy• Stage of development• Exit strategy: Sell asset? At what phase?• Partner with large pharma, biotech?

• Risk• Postpone internal manufacturing costs?• Build internal manufacturing later, if needed?

Page 8: Next Generation Recombinant Protein Manufacturing

CMO Option• Advantages of Using CMO

• Contract vs build (rent vs buy)• Access to existing facilities• Access to CMO expertise

• Selection Criteria• Strong and relevant experience• Scientific expertise• Implementation of latest innovations• Quality systems / regulatory history• Program management

• Geographic location of CMO• Close proximity not essential• Good communications are critical

Page 9: Next Generation Recombinant Protein Manufacturing

Single Use Bioreactor vs Stainless Steel

Page 10: Next Generation Recombinant Protein Manufacturing

SUB vs SS Considerations

Scale of Manufacturing

Quality

Flexibility

SUB vs SS Comparison

Page 11: Next Generation Recombinant Protein Manufacturing

Scale of Manufacturing

*Yields are based on upstream expression levels.with downstream recoveries assumed to be 70%.

Page 12: Next Generation Recombinant Protein Manufacturing

SUB Quality

Single Use Bioreactors:

• No CIP/SIP reduces changeover time.

• Significantly decreases possibility of product carryover or cross-contamination from one campaign to the next.

• Increased facility throughput.• Reduced timelines for sequential

batch manufacturing.• Flexible manufacturing scale

matches process scale to DS requirements, reducing materials costs.

Page 13: Next Generation Recombinant Protein Manufacturing

SUB Flexibility• Flexible manufacturing scale of GE xcellerex SUB with

a 5:1 turndown ratio – 2000L SUB working volume scalable from 400L to 2000L.

• SUB maximum working volume of 2000L could be limitation.

• Multiple DS batches at 2000L may satisfy DS requirements – e.g. 2 X 2000L and possible pooling for purification as single batch.

• SUB shows good comparability to SS such that transfer to larger scale SS bioreactor can be made for late stage / commercial, if needed.

Page 14: Next Generation Recombinant Protein Manufacturing

SUB vs Stainless SteelSingle Use Facility Relative to Stainless Steel

Water Usage 87% reductionCleaning Chemicals Usage 95% reductionEnergy (Electricity) Demand 30% reduction

Facility Footprint 38% lessSteelwork 62% lessHeadcount 21% lower

Plastic Waste 880 kg increaseCO2 Emissions 26% reduction

Comparison based on 3 X 2000L MAb Production Scale*

* The Environmental Impact of Disposable Technologies By Andrew Sinclair, Lindsay Leveen,Miriam Monge,Janice Lim,Stacey Cox

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Accelerated Manufacturing Timelines

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Time is of the Essence

Bring innovative therapies to patients sooner.

Innovative biologic drugs improve patient survival, quality of care, and quality of life.

Intense competition for share of biosimilar market requires early approval and entry into market to be

successful.High cost of drug development requires shortening time

to return on investment for venture capitalists and investors.

Need to exploit any opportunities to shorten clinical trial / regulatory approval process without sacrificing quality.

Page 17: Next Generation Recombinant Protein Manufacturing

Opportunities to AccelerateCell Line Development

Cell Culture Process Development

Purification Process Development

Analytical Method Development

Drug Product Formulation Development

Tox Manufacturing

Page 18: Next Generation Recombinant Protein Manufacturing

Cell Line Development Technologies

Vector Construction

Transfection & Selection

(Amplification)Pool

EnrichmentClone Isolation

& ScreeningStability

Assessment

Strong vector w/ enhance element

Transient Transfection

Gene

Stable Pool

FACS

High throughputcloning

(FACS or ClonePix)

Gene codonoptimization Shaker

24-wp clone

screening

CSI

Process Development

Process Development

Cell Bank

High Throughput clone selection (Clonepix, FACS) screens largernumber of clones and selects clones based on productivitycompared with traditional limited dilution cloning.

Page 19: Next Generation Recombinant Protein Manufacturing

Shaker 24-Well Plate Clone Screening

10 days

14‐21 days

10 days

Static culture

Shaker culture (deep 24‐wp)

10 days

14‐21 days

0

5

10

15

20

25

30

35

40

45

0 5 10 15 20 25 30 35 40Cl

one

Rank

ing

by d

24w

pClone Ranking in SF 7 day batch culture

New cloning screening process reduces timeline by 10 days. Shaker culture is introduced as early as possible, so that the clones screened out fit into the downstream scale up model.

Good correlation between two steps

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High Throughput Cell Culture Process Development

Basal medium, feed medium, and process parameter studiescan be performed at 9-16 ml scale rather than 2L or 10Lworking volume. Conventional process development timelinesof 3-4 months can be reduced to 1-1.5 months using ambr.

1mL tips

1mL or

4mL tips

Two culture stations; each holding 12 bioreactors.All 24 bioreactors independently controlled for pH and DO.

Used Tips Discard

LiquidHandler

ambrTM Technology

Microbioreactor(9-16mL working volume)

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Scale Up Studies

Cell Growth Titers Product Quality Attributes

• Comparison across scales for the production of a recombinant glycoprotein in a recombinant CHO cell line.

• The process decisions and results from ambrTM were reproducible to other scales.

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Platform mAb Purification Process

Bulk Fill

UF/DF: formulation buffer exchange, concentration adjustment

Virus filtration

Chromatography (can be flow through)

Purification IEX chromatography

Low pH viral inactivation

Protein A capture chromatography

Cell separation by depth filtration

Page 23: Next Generation Recombinant Protein Manufacturing

High Throughput Analytics

LabChip GXII has capability to run specific assays for Protein Glycan, Protein Charge Variant (CZE), and Protein Molecular Weight

Octet instrument is used for quick turnaround ProA-based titer analysis of diluted, high concentration, in process cell culture samples.

High Throughput Analytics allow for rapid titer determinations and assessment of critical quality attributes, leading to faster decisions and reduced timelines in cell line development and process development.

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Analytical – Utilize Platform Methods as Appropriate

• Protein Primary Structure Peptide Sequencing via LC/MS/MS Amino Acid Analysis Peptide Mapping

• Biophysical Characterization CD, FTIR, DSC, DLS, fluorescence

spectroscopy

• Capillary and Slab Gel Electrophoresis CZE SDS-CGE cIEF and icIEF SDS-PAGE and IEF Western blot Microchip electrophoresis 2D gels and blots

• Glycan Analysis Oligosaccharide mapping Monosaccharide composition Sialic Acid Quantitation

• Process Residuals• ELISA (HCP, protein A etc.)• HPLC (antibiotics, IPTG, detergents, etc)• qPCR (DNA)

• HPLC• Size Exclusion (with MALLS)• Ion Exchange• Reverse Phase• Hydrophobic Interaction• Affinity

• Potency Assays• Binding Assays via ELISA, Biacore and

ForteBio• Cell Based Assays (e.g., proliferation,

cytokine release, etc.)

• Mass Spectrometry• Intact mass• Peptide mapping with LC/MS or

LC/MS/MS• Disulfide Mapping• Post translational modifications (e.g.,

oxidation, deamidation)• PEGylation site identification• Glycan Identification & site identification

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Preformulation: Design of Experiments (DOE)

30-40 Candidate Formulations:

Various buffer, pH, ionic strength, and excipient conditions

Biophysical Screening: DSC, DLS, CD, FTIR, Fluorescence

Select Candidate formulation(s)

with appropriate thermal and

structural stability

High Throughput Preformulation screening approach selects optimal formulations to carry into forced degradation and final dosage development based on biophysical characteristics and guided by statistical analysis.

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Platform Expansion Process

Wave 20/50

Vial Thaw

Xcellerex 2000L

Xcellerex 200L

Shake Flask

Expansion

Seed or Production Bioreactor

Production Bioreactor

Each passage 2 – 4 daysProduction 2 – 3 weeks

Page 27: Next Generation Recombinant Protein Manufacturing

Production Options for Preclinical Tox Drug Substance Production

Conservative Approach

• Engineering run performed in cGMP suite

• Same single use bioreactor platform as PD

• Scale up to 2000L• Source of reference material• Viral clearance sampling• More control• Less risk• Traditional timeline• Slot reservation considerations

Accelerated Approach

• Demonstration run performed in process development suite

• Same single use bioreactor as cGMP

• Scale limited 200L• Source of reference material• Viral clearance sampling• Less control• More risk• Shorter timeline• More flexibility in scheduling

Page 28: Next Generation Recombinant Protein Manufacturing

Conclusions• Using a CMO for production of early clinical

drug substance offers a compelling value proposition.

• Employing Single Use Bioreactors provides a strong quality advantage and maintains flexibility in terms of clinical and commercial manufacturing.

• Reducing timelines through effective use of technology innovations and manufacturing strategies may shorten time to approval and increase return on investment.

Page 29: Next Generation Recombinant Protein Manufacturing

Thank you!


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